非模型酵母Issatchenkia orientalis SD108的代谢工程用于5-氨基氨酸生产
Shih-I Tan1,2, I-Son Ng2, Huimin Zhao1
1Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana Champaign, Champaign, Illinois, USA.
Biotechnology and bioengineering
|November 7, 2024
概括
这项研究设计了一种耐酸酵母,Issatchenkia orientalis,用于增强5 - 氨基诺氨酸 (5-ALA) 的生物生产. 代谢工程策略显著提高了5-ALA的产量,证明了大规模应用的潜力.
科学领域:
- 生物技术是生物技术.
- 代谢工程是代谢工程.
- 微生物生产 微生物生产
背景情况:
- 人们越来越感兴趣的生物生产的5-aminolevulinic 酸 (5-ALA).
- 在5-ALA生产过程中发酵的酸化会限制细胞生长和产量.
- 鉴定出Issatchenkia orientalis SD108是5-ALA生物合成的有希望的耐酸宿主.
研究的目的:
- 设计一种耐酸酵母,以改善5-ALA的生产.
- 为了克服 5-ALA 发酵中汁酸化的局限性.
- 通过代谢工程来增强5-ALA的微生物生产.
主要方法:
- 设计了耐酸酵母Issatchenkia orientalis SD108.8. 这是一种耐酸酵母.
- 优化了等离子体设计,过度表达了输送器,并增加了5-ALA生产的基因拷贝数.
- 重定向代谢流量,创建一个pyruvate脱碳酶 (PDC) 淘汰菌株 (SD108ΔPDC),并利用尿素作为源.
主要成果:
- 发现5-ALA可以提高I. orientalis SD108.8中的细胞生长率.
- 鉴定了一种内源性ALA合成酶 (ALAS),在I. orientalis.中具有较高的活性.
- 通过代谢工程和优化发酵条件,实现了5-ALA标位的13倍增加,达到510毫克/升.
结论:
- 经过工程改造的耐酸菌株I. orientalis SD108ΔPDC显示出大规模生产5-ALA的巨大潜力.
- 代谢流量重定向和战略源选择对于最大限度地提高5-ALA产量至关重要.
- 鉴定到的IoALAS酶和耐酸性宿主是有效的5-ALA生物合成的关键因素.
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